JPH0458157A - Automatic enzyme immune measuring instrument - Google Patents

Abstract

PURPOSE:To measure a large quantity of samples in a short period of time by successively dispensing the samples into many reagent cartridges inserted to a 1st level to effect an antigen-antibody reaction, dispensing substrates therein to effect enzyme reaction and measuring the reaction. CONSTITUTION:A reagent cartridge stocker 1 lines up and holds the many cartridges 10 in the inserted state within the X-Y plane of the 1st level. The samples are successively dispensed into the many reagent cartridges 10 inserted to this 1st level to effect the antigen-antibody reaction and are further brought into reaction with the labeling antibodies in the cartridges. The enzyme reaction by the injection of the substrates is continuously effected and the measurement is made.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an enzyme immunoassay using magnetic particles, and particularly to an enzyme immunoassay device that is completely automated and can significantly shorten the measurement time.

[Conventional technology]

Enzyme immunoassay (rEIA) has excellent specificity and sensitivity, and is widely used in the field of clinical testing. Normally, the EIA method uses an antigen or antibody depending on the analyte to be measured, bound to a solid phase, which is brought into contact with the analyte in the sample, and then reacts specifically with the enzyme-labeled analyte. The antigen or antibody to be immobilized is reacted with the antigen or antibody to be immobilized (bind), and the unreacted enzyme-labeled antibody (free) is completely removed by repeating washing (bind/free separation; hereinafter referred to as rB/F separation). After this, the activity of the enzyme labeling the antibody bound to the object to be measured for fatigue is measured,
This method quantitatively measures the target substance in the sample.

Therefore, in carrying out the EIA method, very complicated operations such as dispensing, dilution, stirring, B/F separation, and movement of the solid phase were required.

In order to perform the EIA method with high sensitivity and speed, a method has been reported that uses magnetic particles as the solid phase and performs B/F separation using a special container and a magnetic separation device containing a magnet that fits the container. (See Japanese Patent Application Laid-Open No. 61-273453). Furthermore, partially automated measuring instruments have been developed to measure large amounts of specimens, and various types are known depending on the solid phase and antibody label used ("Enzyme Immunoassay", written by Eiji Ishikawa). , Igaku Shoin, pages 180-207).

[Problem to be solved by the invention]

In immunoassays using such techniques, steps such as suction and separation of reagents and samples, stirring, B/F separation associated with various reactions, washing, and measurement have traditionally been performed individually and mainly manually. Measurement of a large number of samples requires a great deal of effort and time by skilled personnel.

An object of the present invention is to provide an automatic immunoassay device that can completely automate such immunoassays, is relatively small, and is capable of measuring a large amount of samples in a short period of time.

[Failure to solve the problem]

The automatic immunoassay device according to the present invention is at a first level,
means for sequentially moving a plurality of cartridges each having a first well enclosing an enzyme-labeled antibody, a second well without inclusions, and a third well enclosing sensitized magnetic particles to a predetermined position within the level; Lifting means for sequentially transporting the cartridges to be transferred to a second level above it, and a reaction line for receiving one after another in a stepwise manner the cartridges on this second level that are successively lifted by the lifting means;
Move to the individual tip on this second level sampling tip cassette to load it, then move to the sample position on the sample cassette also placed on the second level and aspirate that sample. means for dispensing the diluent into a second well of the cartridge located at a second position on the reaction line; and a means for transferring it to the third well of the cartridge after agitation, and a means disposed along this reaction line to agitate the contents of the third well of the cartridge on the line to generate an antigen-antibody reaction and perform B/F separation. means for dispensing the substrate into a third well and stirring it to cause an enzyme reaction, and an optical measuring device for measuring the reaction a predetermined time after the start of the enzyme reaction. and means for providing operation timing of these elements.

[For production]

Samples are sequentially dispensed into a number of pre-prepared reagent cartridges inserted into the first level to perform antigen-antibody reactions, further react with labeled antibodies in the cartridges, and then undergo enzyme reaction by injecting substrates. Measurement is performed by continuously performing

〔Example〕

FIG. 1 is an external perspective view of the automatic enzyme immunoassay device of the present invention, and FIG. 1a is a longitudinal sectional view of a reagent cartridge used in the device.

In FIG. 1, a reagent cartridge stocker 1 shown by a phantom line in the lower left portion is movable in the direction of the arrow and supports a plurality of sample cartridges 10 in rows and columns. In this embodiment, the reagent cartridge 10 has, as shown in FIG. 1a, a third well prefilled with an enzyme-labeled antibody at the left end, a first well prefilled with a solution containing sensitized magnetic particles at the right end, and It has a substantially rectangular shape with a second well without filling in the center, and has notches at both ends for stable engagement with related elements described below. These wells are sealed by seals 11 as shown in FIG. 1a.

The first cartridge to be inserted into this reagent cartridge stocker 1
Above the level, there is a part 5 which can be opened and closed, including a sample part and a reaction line which will be described later, a pump part 6 which can also be opened and closed,
A chip disposer 7 and a measurement system 8 are arranged on the side thereof, and a cartridge disposer 9 is further provided.

2 is a plan view of the apparatus with the cover of FIG. 1 removed, and FIG. 3 is a side view taken along line nn in FIG. 2.

2 and 3, the reagent cartridge stocker 1 stores a number of cartridges 10 in the inserted state.
They are maintained in matrix in the X-Y plane of the level. A cartridge crane mechanism is disposed above the cartridge stocker 1 so that the cartridges on the stocker 1 can be sequentially picked up and transferred to a predetermined position. On a second level above this cartridge crane mechanism, the sampler partial reaction line, measurement system, and various elements related to the reaction, which form the main part of the device, are arranged, and above this, a sample crane mechanism similar to that described above is arranged. is located. The configuration requirements of this device will be explained in detail for each level below.

As described above, the cartridge stocker 1 that holds the reagent cartridge 10 can be inserted into and taken out from the apparatus in the horizontal direction.

Cartridge Crane Mechanism As shown in FIGS. 2 and 3, a cartridge crane mechanism is arranged above the cartridge stocker 1. As shown in FIGS. This mechanism consists of a pair of rails 2 extending parallel to the insertion direction of the cartridge stocker 1 and perpendicular to that direction.
1 includes a crane arm 20 movable along 1. A cartridge pickup device 22 is mounted on the arm 20 so as to be movable along the arm 20.

The mechanism for realizing the movement of the arm 20 along the rail 21 and the movement of the pickup device 22 along the arm 20 is well known in the X-Y block, etc., and in this embodiment, it is driven by a reversible motor (not shown). This is achieved by connecting the arm 20 or the pickup device 22 to a wire placed around a pulley (not shown).

It is clear that other mechanisms may be used for this purpose.

These reversible motors are controlled so that the pickup device 22 sequentially transports the cartridges 10 aligned on the cartridge stocker 1 one by one to predetermined positions (indicated by A in FIG. 2) at timings to be described later.

Pickup Device Pickup device 22 is shown in FIGS. 4A-40. This pickup device 22 has a vertically movable hook device,
Figure 4A shows the hook device in the upper position;
Figure B is a front view showing a state in which the hook device has descended and gripped the cartridge 10 located below;
The figure is a side sectional view of FIG. 4A. In these drawings,
The pickup device 22 is carried onto a predetermined cartridge in the state shown in FIG. 4A. At that position, the hook device is lowered to grip the cartridge, and then returned to the state shown in FIG. 4A and transported to position A in FIG. 2, where the hook device is lowered again to separate the gripped cartridge. It works like this.

The pickup device 22 has a substantially inverted U-shaped frame 221 that is slidably engaged with the arm 20.
21 has a pair of vertical guide rods 222 between the legs. Slider members 223 are slidably attached to these rods 222. As shown in FIG. 4C, a horizontal groove 224 is formed on the back of the slider member 223.
A motor 224 is attached to the back of the frame 221. An arm 225 is fixed to the shaft of this motor 224, and a slider member 22 is attached to the other end of this arm 225.
a protrusion 226 that slidably fits into the transverse groove 224 of 3;
is formed. Therefore, the protrusion 226 slides within the lateral groove 224 as the motor 224 rotates, thereby causing the slider member 223 to move up and down.

A plate 227 is attached to the front of the slider member 223 for the bin 2.
28 at intervals. A pair of cam levers 22 are provided between the slider member 223 and the plate 227.
9 is arranged to rotate around the pin 228.

A pair of hook members 230 are attached to the front surface of the plate 227 so as to rotate around the bin 231. A hook 230 is formed at the lower end of this hook member 230, and the other end is provided in the plate 227 and the opening 23
3 and is rotatably connected to the tip of the cam lever 229 by a pin 232.

These hook members 230 are attached to the spring 2 as shown in FIG. 4A.
34, and therefore, for example, looking at the right cam lever 229 in the figure, its tip is biased counterclockwise. In order to maintain the state of the hook member 230 shown in FIG. 4A, the clockwise movement of the cam lever 229 is restricted by a stopper 235.

When the motor 224 rotates and the slider member 223 descends, the hook member 230 also descends in the state shown in FIG. 4A. When the lower end of the cam lever 229 hits the lower edge of the frame 221 and further descends, the lower end of the cam lever 229 is rotated clockwise by the lower edge. Therefore, the hook member 2
When the upper end of hook 30 rotates counterclockwise, hook 230a rotates counterclockwise against spring 234.

For the configuration on the left, the opposite occurs, resulting in the hooks 230a moving apart from each other. Because the top of the cartridge is positioned in this vertical position,
Open hook 230a grips the cartridge as shown in Figure 4B. As the slider member 223 is subsequently raised, the action of the lower edge of the frame 221 is eliminated, but the grip of the cartridge by the hook 230a is maintained by the spring 234.

In this state, the pickup device 22 is moved to position A in FIG. 2, lowers the slider member 223 again, engages the lower end of the cam lever 229 with the lower edge of the frame 221, opens the hook 230a again, and moves the cartridge to that position. Complete the transfer. The pickup device then returns to the state shown in FIG. 4A and is moved to the next cartridge position.

Lift Mechanism The lift mechanism 30 is a mechanism for moving the cartridge 10 that has been moved to position A in FIG. 2 to the reaction line on the second level, and is located adjacent to position A as shown in FIG. .

The lift mechanism 30 will be explained with reference to FIG. 5A and FIG. 5B, which shows a cross section taken along line VI-Vl in FIG. 5A. 5A and 5B, a pair of vertically disposed sprockets located within a frame 306 and driven by a reversible motor 305 (FIG. 5B), one of which is secured to the outside of the frame 306. 301 has chain 302
The rear end of the arm 304 is attached to a point 310 on the left side of the chain 302 in FIG. 6A. This arm 304 is horizontally movably supported by a horizontal slot 311 provided in a holder 305 which is vertically movable within a frame 306 by movement along the chain 302 due to its coupling with the chain 302. There is.

The front part of this arm 304 passes through a vertical slot 307 provided in the front face of the frame 306, and its vertical movement is guided by this slot 307. A pickup device 22' similar to the pickup device shown in FIG. 4 is attached to the front end of the arm 304.

The reversible motor 305 has a connecting point 310 between the chain 302 and the rear end of the arm 304, and a point where the chain 302 intersects the center line of the horizontal arm 304 passing through the axis of the lower sprocket 301 as shown in FIG. 5A (left side). )B to a similar point (right side) C where the upper half of the upper sprocket 301 is filled.

That is, in FIG. 5A, the arm 304 is in the most retracted state, and by the movement of the chain 302 in the direction of the arrow, it rises to the position of the upper sprocket while maintaining this state, and is then connected by a half circumference above the upper sprocket. The position C becomes the most pushed out state due to half the circumference of the point 310. Therefore, the pickup device 22' attached to the tip of the arm 304 traces a trajectory shown by a dotted line. In particular, the arcuate locus in the process of protruding the arm 304 corresponds to the reaction line 70 disposed at the second level.
This is important for stable placement of the cartridge.

To grip and release the cartridge of the pickup device 22', the slider member is lowered to grip the cartridge at the lower arm position in FIG.
2' and lower the slider member again at the upper arm position in FIG. 5A to release the cartridge. These operations are as described in FIG. 4.

(2) Configuration of Second Level Sampler Section As shown in FIG. 2, a sampling chip cassette 50 and a sample cassette 60 are arranged at the second level. A plurality of sampling chips 61 are arranged in a plane in the sampling chip cassette 50, and a plurality of sample containers each containing a specimen are arranged in a plane in the sampling cassette 60.

Reaction Line As also shown in FIG. 2, a reaction line extends in a direction perpendicular to the insertion direction of the first level cartridge stocker 1.

As shown in FIGS. 2 and 3, this reaction line consists of a pair of endless belts 71 arranged in parallel at a distance from each other, and has a structure in which both ends of the cartridge 10 are supported by both belts 71.

The belt 71 is stepped, for example, at intervals of 30 seconds by sprockets 73 driven by step motors 72 provided at both ends.

The cartridge 10 carried from the first level by the lift device 30 in FIG. Repeat. This reaction line is equipped with various mechanisms, which will be described later.

Seal Removal Mechanism A seal removal mechanism 40 for removing the seal 11 of the cartridge 10 lowered to the left end position of the reaction line 70 by the lift device 30 in FIG. 5 is provided at the next position.

In this embodiment, a means is used to make a hole in the seal 11 by lowering a tool having three protrusions on the lower surface. Details of its structure are omitted here.

As shown in Figure 2.3, a mechanism similar to the first level cartridge crane arm mechanism is provided on top of the second level requirements above. This mechanism consists of a pair of rails 80 and an arm 81 movable along them, and unlike the first level mechanism, a sampling pump unit 82 is mounted movably along the arm 81. The two-dimensional operation of this sampling pump unit 82 is obtained in a manner similar to that of the pickup device 22 at the first level. Here, the sampling pump unit 82 will be explained in detail.

Sampling Pump Unit The sampling pump unit 82, shown in front view in FIG.
connected by. The nozzle part 821 is a nozzle 823
The nozzle 823 is fixed on the plate 825 and can be moved vertically by means of suitable guide means.

Any means for providing this vertical movement may be used, such as using a solenoid device. In the lowered state, the nozzle 823 engages the tip 61 of the second level sampling tip cassette 50.

The pump section 822 for performing suction and dispensing functions through the nozzle 823 can take the form of a syringe driven by a solenoid.

A measurement system 90 is provided downstream of the optical measurement system reaction line 70 . The measurement system includes a photomultiplier tube, etc., and the details will be described in the explanation of the operation of this device.

Other components Along the reaction line 70, various pump means, stirring means, B
/F separation means etc. are arranged. These will also be explained in detail in the explanation of the operation of this device.

■ Position A of the cartridge 10 in Figure 2 at the first level of operation of this device.
The transfer from position A on the first level to the reaction line 70 on the second level and the arrangement are as described above, so details will be omitted. Regarding the subsequent operations, the operations at the moving positions of the cartridge 10 indicated by numbers (1) to (66) above the reaction line 70 shown in FIG. 7 will be described. Note that, as described above, this reaction line 70 advances in units of 30 seconds.

In FIG. 7, the cartridge 10 is lowered by the lift mechanism 30 to position (1) in the reaction line 70. After 30 seconds, the cartridge 10 reaches position (2), and the seal removal device 40 (FIG. 2) placed at that position descends to punch a hole in the seal 11 of the cartridge 10 and remove the three wells. Open it.

Next, at position (3), the diluent is dispensed into the unfilled middle second well of the cartridge 10 by the diluent dispensing pump 75 through a suitable nozzle. During this time, the second level sampler crane mechanism is actuated to move the sampling pump unit 82 of FIG. 6 into the sample chip cassette 50, e.g., to position a of FIG. Chip 5 in
1 is attached to the tip of the tip, and then, in that state, the tip is moved to the position shown in FIG. 3, further lowered, and the sample 61 below is sucked into the tip 51, and then returned to the intermediate position.

When the cartridge 10 is in the next position (4), the sampling pump unit in position C of FIG. 3 is moved to position C of FIG. This position C corresponds to the position of the middle second well of the cartridge 10 at position (4) in FIG. Here, the sample aspirated into the chip 51 is dispensed into the second well. After stirring the thus diluted sample by re-aspirating and re-dispensing it into the chip 51, it is transferred to the third well containing the sensitized magnetic particles of the cartridge 10 using the same chip 51. agitate it by re-aspirating and re-dispensing. The sampling pump unit is then moved to position d (second
), the chips are dropped into the chip disposer 7 by retracting the nozzle, and the unit is returned to its position for the next operation.

Next, the cartridge is moved to position (5), and between then and position (24), an antigen-antibody reaction is performed at about 37° C. for 10 minutes.

B/F separation is then performed at position (25). For this B/F separation, a B/F separation device is disposed at this position of the reaction line 70 as shown in cross section in FIG. This device consists of a permanent magnet 90 and a member 91 that holds it on one side between belts 71, and a gap between the other belt 71 and this permanent magnet 90 is provided with a first well in which a reaction of the cartridge 10 occurs. is designed to fit inside. Due to the magnetic field created by the permanent magnet 90, the sensitized magnetic particles in which the antigen-antibody reaction has occurred are attracted to the wall of the first well and separated from the unreacted solution. Then, at the next position (26) within the magnetic field, the washing liquid is dispensed into this first well via a suitable tube by the washing liquid dispensing pump 76 (Fig. 2), and the suction and dispensing suction is repeated. to clean particles adhering to the walls.

Next, the cartridge 10 is moved to position (27), where the enzyme-labeled antibody in the third well of the cartridge 10 is aspirated by the reagent suction separation pump 77 (FIG. 2) and dispensed into the first well.

At this position (27), a ninth
A stirring device as shown in the figure is arranged. This device consists of a bar 1 whose one end is attached slightly eccentrically to the axis of the motor 100.
01, with a U-shaped member 102 at the tip of the bar 101.
is formed, and the middle part of the bar is supported by a support made of, for example, an elastic material. This U-shaped member 1
02 is adapted to receive the first well of the cartridge 10 which has been moved from position (26). That is, the bar 101 vibrates due to the rotation of the motor 100, and this vibration
The signal is transmitted to the first well of the cartridge 10 via the character member 102, and the labeled antibody and the antigen-antibody reactant are stirred.

The cartridge is then moved to position (28) and from this position to position (47) for 10 minutes, the reaction temperature is 37°C.
A labeled antibody reaction is performed in .

Cartridge 10 is then moved to position (48). At position (48), a B/F separation similar to that at position (25) takes place, at the next position (49), which is within the magnetic field, a washing similar to that at position (26), and at position (50) Stirring similar to position (27) is performed, B/F separation is performed again at position (51), and its washing is performed at position (52) within the magnetic field. Although each of the above-mentioned devices can be used independently in each of the positions (48) to (52), due to continuity of operation, the B/F separator and stirring device of FIGS. 8 and 9 can be used separately. It is advantageous in terms of space and cost to integrate the parts, an example of which is shown in FIG. In FIG. 10, permanent magnets 90a and 9b integrated by a support 91' are used for B/F separation at positions (48) and (51), and permanent magnets 90a and 9b are used for B/F separation at positions (50) and (51).
53), a modification of the device of FIG. 9 is shown as the stirring device. The positions and actions of the permanent magnets 90a, 9b on the cartridge are the same as in position (25), respectively.

In the stirring device shown in FIG. 10, the tip of a bar 101' eccentrically mounted on a motor (not shown) is supported by a support 103' made of an elastic material, and a transmission member 104 is fixed to the middle part of the bar 101'. A bar 1 with U-shaped members 102a and 102b fixed to the tip from both ends of the member 104
01a and 101b are designed to extend. These U
The relative positional relationship between the character member and the first well of the cartridge is the same as that shown in FIG. 9, and the operation is also almost the same.

When the cartridge 10 is in position (53), the substrate separation pump 78 removes the substrate from the reagent bottle unit 79.
The solution is dispensed into a well, stirred by a stirring device shown in FIG. 8 provided there, and moved to a position (54). Enzyme reaction is carried out at 37° C. for 5 minutes from this position to position (63). During this reaction, optical measurements were taken 1 minute after the start of the reaction at position (56), and at position (64).
The 5th minute measurement is taken at . The apparatus for this optical measurement is shown in FIG.

This measuring device measures the intensity of the color produced by the enzyme bound to the sensitized particles at a specific wavelength, and at the position (56), the light from the first minute after the start of the reaction is measured at an angle of 45 degrees. ) and (64).
, enters the photomultiplier tube 122 via the semi-transparent mirror 121, and its intensity is measured. Also, the measurement at position (64) is performed directly through the semi-transparent mirror 121. If necessary, the position (64
) may also be associated with a revolver. The output of the photomultiplier tube 122 is processed using a CPU having an appropriate program for display, printing, etc.

At position (65), the cartridge is then picked up and dropped into a waste container 99 by a cartridge disposer 98 having a pick-up device and its moving device similar to that of FIG.

The operation timings of the various elements at the first and second levels described above are automated by programs such as a microphone and a processor.

Furthermore, it goes without saying that the antigen-antibody reaction and labeled antibody reaction treatments can be reversed.

Furthermore, the above description of the operation of the present apparatus is for the case where an enzyme immunoassay using an antibody detection system is performed. In addition, this apparatus can perform enzyme immunoassays using a one-step method, a two-step method, a delayed one-step method, and the like.

The operation of this device in the one-step method does not include dispensing the diluent at position (3), B/F separation at position (25), and dispensing labeled antibody at position (27). except that
The operation is similar to that in the antibody detection system described above. At this time, the sample is dispensed into the third well at position (4), and the enzyme-labeled antibody is dispensed into the third well using the same sampling pump unit when dispensing this sample.

Next, the operation of this device in the two-step method is as follows:
) The operation is performed in the same manner as in the antibody detection system described above, except that the diluent is not dispensed in step ). However, it differs from the antibody detection system in that the sample is dispensed directly into the third well at position (4).

The operation of this device in the delay one-step method is based on the position (
The operation is similar to that of the antibody detection system described above, except that the dispensing of the diluent in step 3) and the B/F separation in position (25) are not performed. However, the second
After dispensing the sample into the well, the enzyme-labeled antibody in the first well is dispensed therein, and at position (27) the sample to which the enzyme-labeled antibody in the second well has been dispensed is dispensed into the third well. This is different from an antibody detection system.

〔Effect of the invention〕

According to the present invention, the measurement of enzyme immunoreaction, which conventionally required most of the manual work, is completely automated, and the time required for measurement is also improved by about 50% compared to the conventional method. The automatic immunoassay device of the present invention, which has a two-level configuration, occupies a small area and is advantageous for installation.

[Brief explanation of the drawing]

Fig. 1 is a perspective view showing the external appearance of the device of the present invention, Fig. 1a is a cross-sectional view of a cartridge used in the device, Fig. 2 is a plan view of its interior, and Fig. 3 is a line drawn along the line ■-■ in Fig. 2. FIGS. 4A to 4C are views showing a cartridge pickup device used in this device, FIGS. 5A and 5B are views showing a lift device for raising the cartridge from the first level to the second level, and FIG. Figure 7 shows the sampling pump unit used for sampling at the second level.
The figure shows each position on the reaction line at the second level, Figure 8 is a cross-sectional view showing the B/F separation device, Figure 9 is a cross-sectional view showing the stirring device, and Figure 10 is the figure 8 and 9. FIG. 11 is a diagram showing an optical measurement device. 1... Reagent cartridge stocker 5... Openable/closeable portion 6... Openable/closeable pump section 7... Chip disposer 8... Measurement system 9... Cartridge disposer patent applicant Fujirebio Co., Ltd. Patent Attorney Sumiko Shimozaka” Ishizu No. 77

Claims (3)

[Claims] (1) At the first level, a number of rectangular cartridges each having a row of a first well filled with an enzyme-labeled antibody, a second well without inclusions, and a third well filled with sensitized magnetic particles are sequentially installed. transport means for moving the cartridges to a predetermined position within the level; lifting means for sequentially transporting the cartridges transferred to the predetermined position to a second level above; and lift means for transporting the cartridges sequentially lifted by the lift means at the second level. in a first position, a reaction line which receives one after another in steps at predetermined intervals; a seal removal means which descends into a cartridge disposed in the reaction line and punctures the seal of the cartridge; selectively actuated diluting means for diluting the sample by dispensing a diluent into a second well of the cartridge located at a second position on the reaction line; and a sampling chip located at this second level. It moves to an individual chip on the cassette and loads it, then moves to the sample position on the sample cassette also located on the second level and aspirates the sample to the cartridge in the second position on the reaction line. After dispensing into the second or third well of
If desired, a first dispensing means for dispensing the enzyme-labeled antibody in the first well of the cartridge to a second or third well, or a sample diluted in the second well to a third well; The third on the line is placed
means for agitating the contents of the second or third well of the cartridge at the position; and means for stirring the contents of the second or third well of the cartridge at the reaction line during a predetermined step from the third position of the cartridge on the reaction line. a means for maintaining the temperature at a temperature that promotes the antigen-antibody reaction and the labeled antibody reaction; and a magnetic field is applied to the third well of the cartridge at the fourth position after the predetermined step to separate the bound and free substances and to maintain the separation state. selectively actuated means for washing bound and removing free, and in a fifth position the sample injected with the enzyme-labeled antibody in the first well or the enzyme-labeled antibody in the second well of said cartridge; selectively actuated means for transferring and agitating the well, and means for maintaining the third well of the cartridge at a temperature that produces a labeled antibody reaction during scheduled steps from this fifth position; means for separating bound and free by applying a magnetic field to the third well at a sixth position after the scheduled step, and washing the bound and removing the free;
At the seventh position, a means for dispensing the substrate into the third well and stirring it to cause an enzyme reaction, an optical measurement device for measuring the reaction after a predetermined time from the start of the enzyme reaction, and a cartridge that has completed the measurement for the reaction. In an automatic enzyme immunoassay device, the reaction line is comprised of a pair of endless belt conveyors juxtaposed at a distance approximately equal to the length of the well row portion of the cartridge; Each belt conveyor has a mechanism that is periodically and intermittently driven by a pair of pulleys to sequentially advance the cartridges placed there.
The automatic enzyme immunoassay device is characterized in that the first, second and third stirring means, first and second heat retention means, and first and second B/F cleaning means are arranged between the endless belt conveyor. . (2) Each of the first, second, and third stirring means includes a member mounted slightly eccentrically on the motor shaft, a rod having one end fixed to the member, and a rod fixed to the other end of the rod. 2. The automatic enzyme immunoassay device according to claim 1, comprising a U-shaped member, and wherein the third well of the cartridge transported by the endless belt can pass through the U-shaped member. (3) The automatic enzyme immunoassay apparatus according to claim 1, wherein each of the B/F cleaning means comprises a permanent magnet arranged between the endless belts.